Acta Agriculturae Boreali-Sinica ›› 2022, Vol. 37 ›› Issue (2): 28-34. doi: 10.7668/hbnxb.20192653

Special Issue: Wheat Drought and water saving Biotechnology

• Crop Genetics & Breeding·Germplasm Resources·Biotechnology • Previous Articles     Next Articles

Cloning and Expression Analysis of TaSPP Gene in Wheat

JING Fanli1,2, ZHANG Peipei1, MIAO Yongping2, CHEN Tao2, LIU Yuan2, YANG Delong1,2   

  1. 1.Gansu Provincial Key Laboratory of Airdland Crop Science,Lanzhou 730070,China
    2.College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China
  • Received:2021-11-01 Published:2022-04-28

小麦TaSPP基因的克隆及表达分析

景凡丽1,2, 张沛沛1, 苗永平2, 陈涛2, 刘媛2, 杨德龙1,2   

  1. 1.甘肃省干旱生境作物学重点实验室,甘肃 兰州 730070
    2.甘肃农业大学 生命科学技术学院,甘肃 兰州 730070
  • 作者简介:
    作者简介:景凡丽(1995—),女,甘肃酒泉人,在读硕士,主要从事生物化学与分子生物学研究。
  • 基金资助:
    中央引导地方科技发展专项子项目(ZCYD-2020-2-2)

Abstract:

In order to clarify the expression characteristics and biological functions of the sucrose phosphate phosphatase gene(SPP),and further understand the regulatory mechanism of SPP involved in sucrose biosynthesis.Herein,three TaSPP homologs,TaSPP-5A,TaSPP-5B and TaSPP-5D,located on the fifth homologous group,were cloned using the cDNA of wheat variety Jinmai 47 as the template.The physical and chemical properties,gene structure,cis-acting elements,phylogenetic tree and protein conserved domains of TaSPP were analyzed by the method of bioinformatics.The expression pattern of TaSPP was analyzed by qRT-PCR.The results showed that TaSPP-5A,TaSPP-5B and TaSPP-5D contained eight exons and seven introns.TaSPP-5A and TaSPP-5D encoded 422 amino acids,while TaSPP-5B encoded 413 amino acids.Phylogenetic analysis showed that TaSPP in wheat and its related species belonged to the same evolutionary branch with highly genetic similarity.The specific expression analysis showed that TaSPP genes were expressed in roots,stems,flag leaves,leaf sheaths,flower and seeds,whereas the higher expression levels were identified in flag leaves and stems.The expressions of TaSPP gene could be induced by ABA,PEG-6000,NaCl and IAA,indicating that TaSPP genes could play an essential role in abiotic stress tolerance in wheat.

Key words: Wheat, SPP, Cloning, Abiotic stress, Expression analysis

摘要:

为明确磷酸蔗糖磷酸酶基因(SPP)的表达特征和生物学功能,进一步了解SPP参与蔗糖生物合成的调控机制。以小麦抗旱品种晋麦47的cDNA为模板克隆出3个位于第5同源群上的TaSPP同源基因,分别命名为TaSPP-5ATaSPP-5BTaSPP-5D。并利用生物信息学对小麦TaSPP的理化性质、基因结构、顺式作用元件、系统进化树和蛋白保守结构域等进行分析,通过qRT-PCR分析基因TaSPP的表达模式。结果表明,TaSPP-5ATaSPP-5BTaSPP-5D都包含8个外显子和7个内含子,TaSPP-5ATaSPP-5D编码422个氨基酸,TaSPP-5B编码413个氨基酸。系统进化分析显示,小麦TaSPP基因与小麦的近缘物种在进化上处于同一分支,相似度较高。qRT-PCR表达分析结果显示,TaSPP基因在根、茎秆、旗叶、叶鞘、颖花、种子中均表达,其中在旗叶和茎秆中表达量较高;ABA、PEG-6000、NaCl和IAA胁迫均能诱导TaSPP基因的表达,表明小麦TaSPP基因可能在逆境胁迫过程中发挥重要作用。

关键词: 小麦, 磷酸蔗糖磷酸酶基因, 克隆, 非生物胁迫, 表达分析